Department of Molecular Biosciences, Northwestern University, Evanston, United States.
Elife. 2022 Feb 11;11:e72872. doi: 10.7554/eLife.72872.
While centrosomes organize spindle poles during mitosis, oocyte meiosis can occur in their absence. Spindles in human oocytes frequently fail to maintain bipolarity and consequently undergo chromosome segregation errors, making it important to understand the mechanisms that promote acentrosomal spindle stability. To this end, we have optimized the auxin-inducible degron system in to remove the factors from pre-formed oocyte spindles within minutes and assess the effects on spindle structure. This approach revealed that dynein is required to maintain the integrity of acentrosomal poles; removal of dynein from bipolar spindles caused pole splaying, and when coupled with a monopolar spindle induced by depletion of the kinesin-12 motor KLP-18, dynein depletion led to a complete dissolution of the monopole. Surprisingly, we went on to discover that following monopole disruption, individual chromosomes were able to reorganize local microtubules and re-establish a miniature bipolar spindle that mediated chromosome segregation. This revealed the existence of redundant microtubule sorting forces that are undetectable when KLP-18 and dynein are active. We found that the kinesin-5 family motor BMK-1 provides this force, uncovering the first evidence that kinesin-5 contributes to meiotic spindle organization. Altogether, our studies have revealed how multiple motors are working synchronously to establish and maintain bipolarity in the absence of centrosomes.
虽然中心体在有丝分裂期间组织纺锤体极,但卵母细胞减数分裂可以在没有中心体的情况下发生。人类卵母细胞的纺锤体经常无法维持双极性,因此会发生染色体分离错误,因此了解促进无中心体纺锤体稳定性的机制非常重要。为此,我们优化了在中的生长素诱导的降解结构域系统,以便在数分钟内从预先形成的卵母细胞纺锤体中去除这些因子,并评估其对纺锤体结构的影响。这种方法表明,动力蛋白对于维持无中心体极的完整性是必需的;从双极纺锤体中去除动力蛋白会导致极张开,如果与由动力蛋白-12 马达 KLP-18 耗竭诱导的单极纺锤体结合,动力蛋白耗竭会导致单极完全溶解。令人惊讶的是,我们继续发现,在单极破坏后,单个染色体能够重新组织局部微管并重新建立介导染色体分离的微型双极纺锤体。这揭示了存在冗余的微管分拣力,当 KLP-18 和动力蛋白活跃时,这些力是无法检测到的。我们发现,驱动蛋白-5 家族马达 BMK-1 提供了这种力,这首次表明驱动蛋白-5 有助于减数分裂纺锤体的组织。总之,我们的研究揭示了在没有中心体的情况下,多个马达如何协同工作以建立和维持双极性。